【作者】 仝保慧；

【导师】 关铭； 再帕尔·阿不力孜；

【作者基本信息】 吉林大学 ， 药物分析， 2009， 硕士

【摘要】 本文对来源于新疆特有具有抗老年痴呆和抗血栓活性的民族药用植物草棉活性提取物AB-8-2中异槲皮苷(isoquercetrin)、槲皮黄苷(quercimeritrin)、槲皮素-3’-葡萄糖苷(quercetin-3’-glucoside)、8-甲氧基-山奈酚-7-鼠李糖苷(8-methoxyl-kaempferol-7-rhamnoside)、紫云英苷(astragalin)、山奈酚-3-(6"-对-香豆酰)-葡萄糖苷(kaempferol-3-(6"-P-coumaroyl)-glucoside)、芦丁(rutin)、槲皮素(quercetin)、山奈酚(kaempferol)等采用HPLC-MS~n技术,开展以多反应监测(MRM)检测模式的9组分同时定量研究。同时运用该方法成功的对3个不同批次的AB-8-2样品进行了9组分定量,显示了良好的重现性和准确度。MS~E或IDA是通过合理地设置质谱的扫描方式可以迅速获得生物样品中被分析测试物的多维信息的LC-MS/MS技术。根据大多数药物的代谢产物基本保持了原药的骨架特征抑或是亚骨架特征,代谢物的质谱裂解行为与原药相近,综合运用LC-MS/MS技术可迅速找到可能的代谢物,并进行结构鉴定的这一理论,初步探讨了大鼠灌胃该活性提取物后的尿液样品与胆汁样品中的代谢产物进行IDA与MS~E代谢产物分析,并对其代谢产物进行了这两种代谢产物分析方法的方法学比较研究。为整体研究中药的代谢及其辩证的作用机制作基础性研究,另外,通过分析这种高通量式的代谢产物信息获取及分析技术,为研究疾病本身提供理论依据。更多还原

【Abstract】 For the modernization of traditional Chinese herbal medicine, quanlity control and the establishment of dependenble pharmacological assessment system for the crude materials and products are considered as the two principal challenges. And at present, it is not enough to evaluate the quality of TCM using one single component. Simultaneous quantitation of multi-components on the basis of pharmacological study has attracted the interests of researchers in the quanlity control of the TCM.AB-8-2 was an active fraction obtained from Gossypium herbaceam L, which mainly consisted of flavonol glycosides. In this study, a method using liquid chromatography coupled to tandem mass spectrometry （LC-MS/MS） for the simultaneously determination of the 9 major constituents of this active fraction. And this method was successfully applied to the quantification of nine constituents in 3 AB-8-2 samples. Data acquisition under MS/MS was achieved by applying multiple reaction monitoring （MRM） of two fragment ions transitions to provide a high degree of sensitivity and specificity for both quantification and confirmation. Nine constituents are rutin, isoquercetrin, quercimeritrin, quercetin-3’-glucoside, 8-methoxyl-kaempferol-7-rhamnoside, astragalin, kaempferol-3-（6"-P-coumaroyl）-glucoside, quercetin, kaempferol eta.The metabolisms in bile and urine could reflect the whole profiles of biotransformation of the parent medicine or compounds. In this paper, the metabolites of flavonols in the bile and urine samples after oral administistration of AB-8-2 were analyzed using LC-MS/MS in a single chromatographic run. Data were aquried using RRLC^TM coupled with a QTOF mass spectrometer. The Information Depended Acquirer （IDA） and MS^E （where E represents collision energy） data acquisition methods are compared and it was shown that the latter has been provide to be a powerful approach for the rapid metabolites identification.Based on the IDA method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 38 constituents including 26 glucuronidated metabolites, 5 sulfation metabolites, 4 methylation metabolites , 3 flavonol aglycones and 1 glucuronidated conjudge methylation metabolite were characterized. Flavonol diglycosides in AB-8-2 probably remained one glycoside in the structure during the absorption process in intestinal tract. In bile sample, 3 constituents including 1 glucuronidated metabolite, 2 sulfation metabolites were characterized.Based on the MS^E method established, the urine sample after oral administration of AB-8-2 was analyzed. In one analytical run, 34 constituents including 25 glucuronidated metabolites, 4 sulfation metabolites, 3 methylation metabolites , 4 flavonol aglycones and 1 glucuronidated conjudge sulfation metabolite were characterized. In bile sample, 15 constituents including 13 glucuronidated metabolites, 1 flavonol aglycones metabolites and 1 methylation metabolite were characterized.In comparision with MS^E, IDA is an artificial intelligence-based produce ion scan mode providing automatic“on-the-fly”MS to MS/MS switching. And the fragmentation spectrometry was more characteristic. However, MS^E produced the same product ion profile but more comprehensive and overall. Although since no precursor-ion selection occurs in the mass spectrometer good chromatographic separation is a critical factor in affording fragment ions that are derived predominantly from the analyte of interest, MS^E enables the almost simultaneous acquisition of both LC-MS and fragmentation data from a single experiment. Conversely, IDA has its inherent drawback that has the capacity to generate some non-compound-related MS/MS spectra or miss potential metabolites.更多还原